Genetic Encoding of a Bicyclo[6.1.0]nonyne-Charged Amino Acid Enables Fast Cellular Protein Imaging by Metal-Free Ligation

被引：149

作者：

Borrmann, Annika ^{[1
]}

Milles, Sigrid ^{[2
,3
]}

Plass, Tilman ^{[2
,3
]}

Dommerholt, Jan ^{[1
]}

Verkade, Jorge M. M. ^{[4
]}

Wiessler, Manfred ^{[5
]}

Schultz, Carsten ^{[2
,3
]}

van Hest, Jan C. M. ^{[1
]}

van Delft, Floris L. ^{[1
]}

Lemke, Edward A. ^{[2
,3
]}

机构：

[1] Inst Mol & Mat, NL-6525 AJ Nijmegen, Netherlands

[2] EMBL, Struct & Computat Biol Unit, D-69117 Heidelberg, Germany

[3] EMBL, Cell Biol & Biophys Unit, D-69117 Heidelberg, Germany

[4] SynAffix BV, NL-6525 AJ Nijmegen, Netherlands

[5] DKFZ, D-69120 Heidelberg, Germany

来源：

CHEMBIOCHEM | 2012年 / 13卷 / 14期

关键词：

amber suppression; click chemistry; Diels-Alder reaction; protein engineering; unnatural amino acid; AZIDE-ALKYNE CYCLOADDITION; FREE CLICK CHEMISTRY; INVERSE-ELECTRON-DEMAND; DIELS-ALDER REACTIONS; COPPER-FREE; MAMMALIAN-CELLS; LIVING CELLS; RECOMBINANT PROTEINS; TRANS-CYCLOOCTENES; TERMINAL ALKYNES;

D O I：

10.1002/cbic.201200407

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Visualizing biomolecules by fluorescent tagging is a powerful method for studying their behaviour and function inside cells. We prepared and genetically encoded an unnatural amino acid (UAA) that features a bicyclononyne moiety. This UAA offered exceptional reactivity in strain-promoted azidealkyne cycloadditions. Kinetic measurements revealed that the UAA reacted also remarkably fast in the inverse-electron-demand DielsAlder cycloaddition with tetrazine-conjugated dyes. Genetic encoding of the new UAA inside mammalian cells and its subsequent selective labeling at low dye concentrations demonstrate the usefulness of the new amino acid for future imaging studies.

引用

页码：2094 / 2099

页数：6

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